Identifification and validation of ferroptosis signatures and immune infifiltration characteristics associated with intervertebral disc degeneration

Feng Zhang; Di Cui; Kangkang Wang; Huimin Cheng; Yunlei Zhai; Wei Jiao; Zhaodong Wang; Xilong Cui; Haiyang Yu

doi:10.3389/fgene.2023.1133615

Identifification and validation of ferroptosis signatures and immune infifiltration characteristics associated with intervertebral disc degeneration

Front Genet. 2023 Feb 22:14:1133615. doi: 10.3389/fgene.2023.1133615. eCollection 2023.

Authors

Feng Zhang^{1

2}, Di Cui³, Kangkang Wang^{1

2}, Huimin Cheng³, Yunlei Zhai^{1

2}, Wei Jiao^{1

2}, Zhaodong Wang^{4

5}, Xilong Cui^{1

2}, Haiyang Yu^{1

2}

Affiliations

¹ Department of Orthopedics, Affiliated Fuyang People's Hospital of Anhui Medical University, Fuyang, Anhui, China.
² Clinical Research Center for Spinal Deformity of Anhui Province, Fuyang, Anhui, China.
³ Medical School of Fuyang Normal University, Fuyang, Anhui, China.
⁴ Anhui Province Key Laboratory of Tissue Transplantation, Bengbu Medical College, Bengbu, Anhui, China.
⁵ Department of Orthopedics, the First Affiliated Hospital of Bengbu Medical College, Bengbu, Anhui, China.

Abstract

Ferroptosis and immune infiltration play an important role in the pathogenesis of intervertebral disc degeneration (IDD). However, there is still a lack of comprehensive analysis on the interaction between ferroptosis-related genes (FRGs) and immune microenvironment in IDD patients. Therefore, this study aims to explore the correlation between FRGs characteristics and immune infiltration in the progression of IDD. The expression profiles (GSE56081 and GSE70362) and FRGs were downloaded from the comprehensive gene expression omnibus (GEO) and FerrDb database, respectively, and the differences were analyzed using R. The intersection of IDD related differential genes (DEGs) and FRGs was taken as differentially expressed FRGs (DE-FRGs) and GO and KEGG enrichment analysis was conducted. Then, we used least absolute shrinkage and selection operator (LASSO) regression algorithm and support vector machine (SVM) algorithm to screen feature genes and draw ROC curve judge the diagnostic value of key DE-FRGs. Then CIBERSORT algorithm is used to evaluate the infiltration of immune cells and analyze the correlation between key DE-FRGs and immune infiltration. Based on the analysis results, we conducted single gene GSEA analysis on key DE-FRGs. RT-PCR and immunohistochemistry further verified the clinical value of the results of biochemical analysis and screening. Seven key DE-FRGs were screened, including the upregulated genes NOX4 and PIR, and the downregulated genes TIMM9, ATF3, ENPP2, FADS2 and TFAP2A. Single gene GSEA analysis further elucidates the role of DE-FRGs in IDD associated with ferroptosis. Correlation analysis showed that seven key DE-FRGs were closely related to immune infiltration in the development of IDD. Finally, RT-PCR and immunohistochemical staining showed that NOX4, ENPP2, FADS2 and TFAP2A were statistically significant differences. In this study, we explored the connection between ferroptosis related characteristics and immune infiltration in IDD, and confirmed that NOX4, ENPP2, FADS2, and TFAP2A may become biomarkers and potential therapeutic targets for IDD.

Keywords: ENPP2; NOX4; ferroptosis; immune infifiltration; intervertebral disc degeneration.

Grants and funding

This study was supported by the Open Project Fund of Ministerial Key Laboratory of Bengbu Medical College in 2022 (Project No. AHTT 2022B003) and Fuyang City Health Commission scientific research project in 2021(Project No.FY 2021-037).